D luciferin

Manufactured by Promega

Sourced in United States, France, United Kingdom, Spain, Japan, Germany, Switzerland

D-luciferin is a chemical substrate used in bioluminescence assays. It is a small organic molecule that emits light when combined with the enzyme luciferase. D-luciferin is a core component in various luminescence-based detection and imaging applications.

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Lab products found in correlation

Living image software, by PerkinElmer (17 mentions) Ivis spectrum, by PerkinElmer (11 mentions) Ivis spectrum in vivo imaging system, by PerkinElmer (9 mentions) Living image 4, by PerkinElmer (9 mentions) Ivis lumina 2, by PerkinElmer (9 mentions) Ivis spectrum imaging system, by PerkinElmer (8 mentions) Ivis imaging system, by PerkinElmer (6 mentions) Ivis kinetic, by PerkinElmer (6 mentions) Pgl4.50 luc2 cmv hygro vector, by Promega (5 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (5 mentions) Ivis spectrum system, by PerkinElmer (4 mentions) Ivis 100 imaging system, by PerkinElmer (4 mentions) Lumina lt, by PerkinElmer (4 mentions) Nightowl 2 imaging systems lb983, by Berthold Technologies (4 mentions) Ivis lumina, by PerkinElmer (4 mentions) Matrigel, by BD (4 mentions) Living image, by PerkinElmer (4 mentions) Ivis lumina series 3, by PerkinElmer (4 mentions) D luciferin, by PerkinElmer (4 mentions) Ami ht, by Spectral Instruments Imaging (4 mentions)

Spelling variants of this product

Luciferin (262 citations) D-luciferin potassium salt (34 citations) D-luciferin substrate (17 citations) D-luciferin solution (5 citations) D-luciferin (beetle) monosodium salt (4 citations) D-Luciferin (Beetle Luciferin Potassium Salt; (3 citations) D-luciferin firefly potassium salt (2 citations) VivoGlo D‐luciferin potassium salt (2 citations) ONE-Glo d-luciferin substrate (2 citations) D-Luciferin sodium (2 citations)

450 protocols using d luciferin

Quantifying In Vitro and In Vivo Bioluminescence

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In vitro BLI and BLI of mice under isoflurane gas anesthesia were carried out on an IVIS Spectrum System (CaliperLS; Perkin-Elmer, Hopkinton, MA, USA). Living Image software (version 4.4.17504) was used for quantifying luminescence.
To measure the in vitro BLI signal intensity, both control and transduced cells were equally seeded (60 000 cells per well in a 24-well plate) and measured the next day after 5 min incubation at 37 °C with D-luciferin (Promega BeNeLux, Leiden, The Netherlands; 150 μg/l dissolved in PBS).
For in vivo BLI measurements, D-luciferin (126 mg/kg dissolved in PBS (15 mg/ml)) was injected intraperitoneally and acquisition of consecutive frames was started until maximum signal intensity was reached. Image acquisition numbers and times varied between 10 and 15 frames of 30-60 s each, depending on the optimal acquisition settings in function of the signal intensity intrinsic to the grade of lung metastasis. We measured the photon flux per second (p/s) through a region of interest (2.9 cm × 1.8 cm) covering the lungs.

Marien E., Hillen A., Vanderhoydonc F., Swinnen J.V, & Vande Velde G. (2017). Longitudinal microcomputed tomography-derived biomarkers for lung metastasis detection in a syngeneic mouse model: added value to bioluminescence imaging. Laboratory investigation; a journal of technical methods and pathology, 97(1).

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Bioluminescent Tumor Imaging in Mice

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Two ID8 luciferase expressing clonal cell sublines, ID8-L4 and ID8-L11, were subjected to bioluminescence imaging. Briefly, cells were seeded in 100 μl medium into a 96-well plate and just prior to imaging using an IVIS Spectrum In Vivo Imaging System (Perkin Elmer, Rodgau, Germany), an equal volume of in vivo glow solution (D-Luciferin, Promega) was added to each well. Images were obtained with a 12.8 cm field of view, 2x2 binning factor and an exposure time of 1 second. As recommended by the manufacturer, the auto-exposure setting was used to automatically set the exposure time, f/stop and binning to keep the signal within an optimal range for quantification and to avoid overexposure during image acquisition. Auto-exposure sensitivity settings used for the snapshot image were adjusted to obtain a minimal target count of 3000. Luminescence was measured as total flux (photons per second (P/S)).
In vivo live bioluminescence imaging of tumor burden was performed using an IVIS Spectrum In Vivo Imaging System. Animals were anesthetised with 1.5-2.5% isoflurane inhalation and given an intraperitoneal injection of 165 mg/kg body weight D-Luciferin (Promega; 30 mg/ml sterile saline) 3 min immediately prior to imaging. Mice were imaged in batches of 4. Images were obtained with a 12.8 cm field of view, 4x4 binning factor and an exposure time ranging from 0.5-1 second.

Clark M., Kollara A., Brown T.J, & May T. (2021). A mouse model of neoadjuvant chemotherapy followed by interval cytoreductive surgery indicates impaired efficacy of perioperative cisplatin. Journal of Ovarian Research, 14, 157.

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NF-κB Activity Modulation Assay

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CL1-5-F4/NF-κB-luc2 cells were plated into 96-well (1.5×10⁴/well) overnight and treated with various concentrations of imipramine (0, 50, 100, 150 and 200 μM), erlotinib (10 μM), hEGF (30 ng/ml), rottlerin (20 nM) and Phorbol myristate acetate (PMA) (75 nM) alone or combined with 100 and 150 μM imipramine for 48 hr. After treatment, 100 μl D-luciferin (Promega, Madison, WI, USA) solution (500 μM D-luciferin) was added into each well and incubated for 5 min in the dark at room temperature before image acquisition. NF-κB activation signal was collected for 3 min by IVIS Lumina LT Series and normalized with cell viability.

Yueh P.F., Lee Y.H., Chiang I.T., Chen W.T., Lan K.L., Chen C.H, & Hsu F.T. (2021). Suppression of EGFR/PKC-δ/NF-κB Signaling Associated With Imipramine-Inhibited Progression of Non-Small Cell Lung Cancer. Frontiers in Oncology, 11, 735183.

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Bioluminescence Imaging of Transplanted Cells

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To visualize the transplanted cells, F3 cells were infected with lentivirus to express the luciferase gene using the method described in the section “Lentivirus preparation and infection”. For the acquisition of bioluminescence images, mice were anesthetized with isoflurane inhalation. d-Luciferin (Promega) was diluted to 3 mg/100 µL in saline and 0.6 mg of d-Luciferin was intravenously administered. An IVIS-100 imaging system (Perkin Elmer) was used, and the images were acquired by integrating light for 30 s. The brains were then isolated from mice and imaged using the IVIS-100 imaging system.

Lee E.J., Choi Y., Lee H.J., Hwang D.W, & Lee D.S. (2022). Human neural stem cell-derived extracellular vesicles protect against Parkinson’s disease pathologies. Journal of Nanobiotechnology, 20, 198.

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Bioluminescence Imaging for Metastatic CDX Cells

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In total, 5 × 10⁵ luciferase-expressing CDX-derived cells were grafted by IC injection into NSG mice (Charles River Laboratories). Metastatic progression was monitored once a week by bioluminescence imaging (BLI) evaluation under anesthesia and after i.p. injection of D-luciferin (15 μg/kg; Promega) using IVIS Spectrum imaging (PerkinElmer). To test in vivo drug efficacy, 2 × 10⁶ cells were injected s.c. into NSG mice. Once tumors reached an average volume of 100 mm³, mice were randomized into groups and treated with vehicle DMSO (1%), olaparib (50 mg/kg, i.p. injection, 3 times per week), BYL719 (20 mg/kg, oral gavage, 3 times per week), AZ82 (10 mg/kg, oral gavage, 3 times per week), or a combination of BYL719 and AZ82. S.c. tumor dimensions were measured by caliper. For BLI evaluation, i.p. with D-luciferin (15 μg/kg; Promega) was performed, and mice were scanned using IVIS Spectrum imaging (PerkinElmer) under anesthesia.

Tayoun T., Faugeroux V., Oulhen M., Déas O., Michels J., Brulle-Soumare L., Cairo S., Scoazec J.Y., Marty V., Aberlenc A., Planchard D., Remon J., Ponce S., Besse B., Kannouche P.L., Judde J.G., Pawlikowska P, & Farace F. (2022). Targeting genome integrity dysfunctions impedes metastatic potency in non–small cell lung cancer circulating tumor cell–derived explants. JCI Insight, 7(11), e155804.

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In Vivo Bioluminescence Imaging

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Tumour-bearing tissues were subjected to in vivo bioluminescence imaging using a Living Image® 4.5.4 IVIS Imaging System (Perkin Elmer). For luciferase imaging, D-luciferin (Promega) was injected intraperitoneally before imaging. Quantitative detection of luciferase was performed as follows: regions of interest (ROIs) were drawn to capture detected uorescence, and auto-regions ROIs were used to precisely outline the target region.

Park M., Cho Y.J., Kim B., Ko Y.J., Jang Y., Hyun H., & Lim W. (2020). RANKL Immunisation Inhibits Prostate Cancer Metastasis by Modulating EMT Through A RANKL-Dependent Pathway.

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Suppressing Cancer Cell Proliferation with Anti-ADAM28 Antibodies

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Example 4

Suppressive Effect of Anti-human ADAM28 Antibodies on Cancer Cell Proliferation In Vivo

ADAM28 high expression breast cancer cell line MDA-MB231^ffLuc-cp156that constitutively expresses luciferase was prepared by a lentivirus vector, and transplanted (2×10e6 cells) to a breast subcutaneous tissue of NOD/SCID mouse (Six-week-old male, Charles River Laboratories International Inc, Washington, Mass.). After transplantation, anti-ADAM28 antibody (2 mg/kg/mice) was topically injected 5 times at 2 day intervals, and the suppressive effect on tumor growth was examined. After intraperitoneal administration of D-luciferin (150 mg/Kg) (Promega Co, Madison, Mich.), the luminescence was detected by in vivo imaging system (IVIS)-100 (Xenogen Co., Alameda, Calif.) (FIG. 3). Both 211-14 and 211-12 antibodies suppressed in vivo proliferation of MDA-MB231^ffLuc-cp156.

Example 6

In Example 4, RT-PCR with luciferase specific primers was performed using RNA extracted from each organ of the mouse at 6 weeks after transplantation of cancer cells, the presence or absence of tumor-derived gene expression was confirmed, and the presence or absence of inartificial metastasis was examined (FIG. 5). Both 211-14 and 211-12 antibodies suppressed micrometastasis of cancer cells.

US10556967B2. Anti-ADAM28 antibody for treating cancer (2020-02-11). GENEFRONTIER CORPORATION [JP], KEIO UNIVERSITY [JP]. Inventors: Akira Miyakoshi [JP], Rena Matsumoto [JP], Shizue Katoh [JP], Yuki Hayami [JP], Satsuki Mochizuki [JP], Masayuki Shimoda [JP], Yasunori Okada [JP].

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Bioluminescence Emission Spectra Analysis

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Bioluminescence emission spectra of D-luciferin (Promega, Madison, WI, USA), CycLuc1 (AOBIOUS INC., Gloucester, MA, USA) and AkaLumine-HCl were measured using an ATTO AB-1850 spectrophotometer (ATTO Co. Ltd, Tokyo, Japan). A reaction mixture was prepared by mixing 5 μl of a substrate (100 μM), 5 μl of QuantiLum Recombinant Luciferase solution (1 mg ml⁻¹) (Promega) and 5 μl of potassium phosphate buffer (500 mM, pH 8.0). Luminescence reactions were then initiated by injecting 10 μl of ATP-Mg (200 μM) into the reaction mixture. Bioluminescence emission spectra were measured in 1 nm increments from 400 to 780 nm using 3 min of integration time.

Kuchimaru T., Iwano S., Kiyama M., Mitsumata S., Kadonosono T., Niwa H., Maki S, & Kizaka-Kondoh S. (2016). A luciferin analogue generating near-infrared bioluminescence achieves highly sensitive deep-tissue imaging. Nature Communications, 7, 11856.

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Bioluminescence Imaging of A20 Leukemia Cells

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Luciferase expressing A20 leukemia/lymphoma cells (A20-luc) syngeneic with recipients (H2d) were used for these experiments (35 (link)). Each recipient mouse received 3×10⁵ A20-luc tumor cells intravenously within 24 hours after BMT. Tumor load was assessed regularly in BMT recipient mice using an Ami 1000 Advanced Molecular Imager (Spectral Instruments, Tucson, AZ) live animal imaging system. Five minutes before BLI, mice were placed in an oxygenated isoflurane chamber and administered D-luciferin (Promega, Madison, WI) intraperitoneally. BMT recipient animals were imaged for 5 minutes and tumor load was quantitated by means of Living Image software v2.50 (Caliper Life Sciences).

Li Y., Chen H.L., Bannick N., Henry M., Holm A.N., Metwali A., Urban JF J.r., Rothman P.B., Weiner G.J., Blazar B.R., Elliott D.E, & Ince M.N. (2014). Intestinal Helminths Regulate Lethal Acute Graft Versus Host Disease and Preserve Graft Versus Tumor Effect in Mice. Journal of immunology (Baltimore, Md. : 1950), 194(3), 1011-1020.

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Liver Metastasis from Gastric Cancer

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The mouse-derived gastric cancer cell line MFC was transfected with lentivirus containing Luc-plasmid (Plenti-CMV-EGFR-Linker-Luc-PGK-Puro) purchased from OOBIO Biotechnology Co., Ltd. (Shanghai). Luc-transfected MFC (Luc-MFC) cells were also transfected with Lv-GPAA1 lentivirus to upregulate the expression of GPAA1 or with or Lv-Ctrl lentivirus as the control. MFC-Lv-Ctrl and MFC-Lv-GPAA1 cells were injected into the spleens of C57BL/6 N mice (n = 5/group) at a concentration of 10⁶ cells/mouse to test the potential capability for liver metastasis. Animals were injected with D-luciferin (150 mg; Promega, catalogue no. P1043), subjected to anaesthesia by isoflurane inhalation, and subjected to luciferin emission imaging to measure the diffusion of cancer cells using an IVIS Spectrum (Caliper Life Sciences) every 5 days. The data were quantified by Living Image software, version 4.5.3. Four weeks later, animals were sacrificed, and an immunohistochemical (IHC) analysis of liver tissue was conducted to evaluate the metastatic ability of the cancer cells.

Zhang X.X., Ni B., Li Q., Hu L.P., Jiang S.H., Li R.K., Tian G.A., Zhu L.L., Li J., Zhang X.L., Zhang Y.L., Yang X.M., Yang Q., Wang Y.H., Zhu C.C, & Zhang Z.G. (2019). GPAA1 promotes gastric cancer progression via upregulation of GPI-anchored protein and enhancement of ERBB signalling pathway. Journal of Experimental & Clinical Cancer Research : CR, 38, 214.

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